Abstract
Postoperative pancreatic fistula (POPF), the major driver of morbidity and mortality following pancreatectomy, is caused by an abnormal communication between the pancreatic ductal epithelium and another epithelial surface containing pancreas-derived, enzyme-rich fluid. There is a strong correlation between the amylase content in surgically-placed drains early in the postoperative course and the development of POPF. A simple and cheap method to determine the amylase content from the drain effluent has been eagerly advocated. Here, we developed an amylase optical biosensor, based on a surface plasmon resonance (SPR) plastic optical fiber (POF), metallized with a 60 nm layer of gold and interrogated with white light. The sensor was made specific by coupling it with an anti-amylase antibody. Each surface derivatization step was optimized and studied by XPS, contact angle, and fluorescence. The POF-biosensor was tested for its response to amylase in diluted drain effluents. The volume of sample required was 50 µL and the measurement time was 8 min. The POF-biosensor showed selectivity for amylase, a calibration curve log-linear in the range of 0.8–25.8 U/L and a limit of detection (LOD) of ~0.5 U/L. In preliminary tests, the POF-biosensor allowed for the measurement of the amylase content of diluted surgically-placed drain effluents with an accuracy of >92% with respect to the gold standard. The POF-biosensor allows for reliable measurement and could be implemented to allow for a rapid bedside assessment of amylase value in drains following pancreatectomy.
Highlights
Given the key role played by the amylase levels in detecting Postoperative pancreatic fistula (POPF), we propose the development of a biosensor for fast, point-of-care, cost-effective and quantitative amylase measurements in surgicallyplaced drain effluent
Bovine serum albumin (BSA, A7030), α-lipoic acid (T5625), anti-α-amylase produced in rabbit fractionated antiserum (A8273) (IgG AMY), anti-mouse polyvalent immunoglobulins (G,A,M)−FITC, antibody produced in goat (F1010) (IgG FITC), and all of the powders for buffer solutions were purchased from Sigma-Aldrich srl (Milan, Italy)
We reported on the development of a surface plasmon resonance (SPR) plastic optical fiber (POF)-biosensor (1)
Summary
In 2005, an international panel of pancreatic surgeons agreed to establish an easy-to-apply clinical definition of POPF: an output from surgically or percutaneously-placed drains of any measurable volume of fluid on or after postoperative day 3, with an amylase content greater than 3 times the upper bound of normal serum amylase activity [1]. While a certain diagnosis of POPF can be established from postoperative day 3 and onwards, POPF is reliably predicted by measuring the amylase value from surgically-placed drains on postoperative day 1 [2]. This allows for tailored fast-track recovery pathways in low-risk patients and the implementation of mitigation strategies in high-risk patients [3]. Reliable, and easy to use point-of-care system to measure the levels of amylase in POPF would be of great importance for an early assessment of high-risk patients and for their treatment
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